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Gut microbiota absence and transplantation affect growth and intestinal functions:An investigation in a germ-free pig model

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Animal Nutrition 2021年第2期7卷 295-304页

作者： Hua Zhou Jing Sun Bing Yu Zuohua Liu Hong Chen Jun He Xiangbing Mao Ping Zheng Jie Yu Junqiu Luo Yuheng Luo Hui Yan Liangpeng Ge Daiwen Chen Key Laboratory of Animal Disease-Resistance Nutrition ChengduSichuan 611130China Animal Nutrition Institute Sichuan Agricultural UniversityChengduSichuan 611130China Key Laboratory of Pig Industry Sciences RongchangChongqing 402460China Chongqing Academy of Animal Sciences RongchangChongqing 402460China College of Food Science Sichuan Agricultural UniversityYa'anSichuan 625014China

This study was conducted to investigate host-microbiota interactions and explore the effects of maternal gut microbiota transplantation on the growth and intestinal functions of newborns in a germ-free(GF)pig model.Twelve hysterectomy-derived GF Bama piglets were reared in 6 sterile isolators.Among them,6 were considered as the GF group,and the other 6 were orally inoculated with healthy sow fecal suspension as fecal microbiota transplanted(FMT)group.Another 6 piglets from natural birth were regarded as the conventional(CV)group.The GF and FMT groups were hand-fed with Co60-y-irradiated sterile milk powder,while the CV group was reared by lactating Bama sows.All groups were fed for 21 days.Then,all piglets and then were switched to sterile feed for another 21 days.Results showed that the growth performance,nutrient digestibility,and concentrations of short-chain fatty acids in the GF group decreased(P<0.05).Meanwhile,the serum urea nitrogen concentration and digesta pH values in the GF group increased compared with those in the FMT and CV groups(P<0.05).Compared with the CV group,the GF group demonstrated upregulation in the mRNA expression levels of intestinal barrier function-related genes in the small intestine(P<0.05).In addition,the mRNA abundances of intestinal development and absorption-related genes in the small intestine and colon were higher in the GF group than in the CV and FMT groups(P<0.05).The FMT group exhibited greater growth performance,lipase activity,and nutrient digestibility(P<0.05),higher mRNA expression levels of intestinal development and barrier-related genes in the small intestine(P<0.05),and lower mRNA abundances of pro-inflammatory factor in the colon and jejunum(P<0.05)than the CV group.In conclusion,the absence of gut microbes impaired the growth and nutrient digestibility,and healthy sow gut microbiota transplantation increased the growth and nutrient digestibility and improved the intestinal development and barrier function of newborn piglets,indicating the importance of intestina

关键词： germ-free Host microbiota interaction Maternal gut microbiota transplantation Growth performance Intestinal function Pig model

Colonization of germ-free mice with a mixture of three lactobacillus strains enhances the integrity of gut mucosa and ameliorates allergic sensitization

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Cellular & Molecular Immunology 2016年第2期13卷 251-262页

作者： Hana Kozakova Martin Schwarzer Ludmila Tuckova Dagmar Srutkova Elzbieta Czarnowska Ilona Rosiak Tomas Hudcovic Irma Schabussova Petra Hermanova Zuzana Zakostelska Tamara Aleksandrzak-Piekarczyk Anna Koryszewska-Baginska Helena Tlaskalova-Hogenova Bozena Cukrowska Laboratory of Gnotobiology Institute of Microbiology Academy of Sciences of the Czech Republic Novy Hradek Czech Republic Department of Pathology the Children's Memorial Health Institute Warsaw Poland Institute of Specific Prophylaxis and Tropical Medicine Medical University of Vienna Vienna Austria Institute of Biochemistry and Biophysics Polish Academy of Sciences Warsaw Poland

Increasing numbers of clinical trials and animal experiments have shown that probiotic bacteria are promising tools for allergy prevention. Here, we analyzed the immunomodulatory properties of three selected lactobacillus strains and the impact of their mixture on allergic sensitization to Bet v I using a gnotobiotic mouse model. We showed that Lactobacillus （L.） rhamnosus LOCK0900, L. rhamnosus LOCK0908 and L. casei LOCK0919 are recognized via Toll-like receptor 2 （TLR2） and nucleotide-binding oligomerization domain-containing protein 2 （NOD2） receptors and stimulate bone marrow-derived dendritic cells to produce cytokines in species- and strain-dependent manners. Colonization of germ-free （GF） mice with a mixture of all three strains （Lmix） improved the intestinal barrier by strengthening the apical junctional complexes of enterocytes and restoring the structures of microfilaments extending into the terminal web. Mice colonized with Lmix and sensitized to the Bet v I allergen showed significantly lower levels of allergen-specific IgE, IgG 1 and IgG2a and an elevated total IgA level in the sera and intestinal lavages as well as an increased transforming growth factor （TGF）-β level compared with the sensitized GF mice. Splenocytes and mesenteric lymph node cells from the Lmix-colonized mice showed the significant upregulation of TGF-β after in vitro stimulation with Bet v 1. Our results show that Lmix colonization improved the gut epithelial barrier and reduced allergic sensitization to Bet v 1. Furthermore, these findings were accompanied by the increased production of circulating and secretory IgA and the regulatory cytokine TGF-β. Thus, this mixture of three lactobacillus strains shows potential for use in the prevention of increased gut permeability and the onset of allergies in humans,

关键词： allergic sensitization germ-free intestinal barrier Lactobacillus probiotics

Oral bacteria colonize and compete with gut microbiota in gnotobiotic mice

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International Journal of Oral Science 2019年第3期11卷 192-200页

作者： Bolei Li Yang Ge Lei Cheng Benhua Zeng Jinzhao Yu Xian Peng Jianhua Zhao Wenxia Li Biao Ren Mingyun Li Hong Wei Xuedong Zhou State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases Sichuan UniversityChengduChina Department of Cariology and Endodontics West China Hospital of StomatologySichuan UniversityChengduChina Department of Laboratory Animal Science College of Basic Medical SciencesThird Military Medical UniversityChongqingChina Shanghai Majorbio Bio-pharm Technology Co. LtdShanghaiChina

The oral microbiota is associated with oral diseases and digestive systemic diseases.Nevertheless,the causal relationship between them has not been completely elucidated,and colonisation of the gut by oral bacteria is not clear due to the limitations of existing research models.The aim of this study was to develop a human oral microbiota-associated (HOMA) mouse model and to investigate the ecological invasion into the gut.By transplanting human saliva into germ-free (GF) mice,a HOMA mouse model was first constructed.16S rRNA gene sequencing was used to reveal the biogeography of oral bacteria along the cephalocaudal axis of the digestive tract.In the HOMA mice,84.78% of the detected genus-level taxa were specific to the donor.Principal component analysis (PCA) revealed that the donor oral microbiota clustered with those of the HOMA mice and were distinct from those of specific pathogen-free (SPF) mice.In HOMA mice,OTU counts decreased from the stomach and small intestine to the distal gut.The distal gut was dominated by Streptococcus,Veillonella,Haemophilus,Fusobacterium,Trichococcus and Actinomyces.HOMA mice and human microbiota-associated (HMA) mice along with the GF mice were then cohoused.Microbial communities of cohoused mice clustered together and were significantly separated from those of HOMA mice and HMA mice.The Source Tracker analysis and network analysis revealed more significant ecological invasion from oral bacteria in the small intestines,compared to the distal gut,of cohoused mice.In conclusion,a HOMA mouse model was successfully established.By overcoming the physical and microbial barrier,oral bacteria colonised the gut and profiled the gut microbiota,especially in the small intestine.

关键词： microbiota germ-free component analysis

Short-chain fatty acids can improve lipid and glucose metabolism independently of the pig gut microbiota

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Journal of Animal Science and Biotechnology 2021年第4期12卷 1414-1427页

作者： Hua Zhou Bing Yu Jing Sun Zuohua Liu Hong Chen Liangpeng Ge Daiwen Chen Key Laboratory of Animal Disease-Resistance Nutrition Chengdu 611130SichuanChina Animal Nutrition Institute Sichuan Agricultural UniversityChengdu 611130SichuanChina Key Laboratory of Pig Industry Sciences Rongchang 402460ChongqingChina Chongqing Academy of Animal Sciences Rongchang 402460ChongqingChina College of Food Science Sichuan Agricultural UniversityYa’an 625014SichuanChina

Background:Previous studies have shown that exogenous short-chain fatty acids(SCFAs)introduction attenuated the body fat deposition in conventional mice and pigs.However,limited studies have evaluated the effects of exogenously introduced SCFAs on the lipid and glucose metabolism independently of the gut microbiota.This study was to investigate the effects of exogenous introduction of SCFAs on the lipid and glucose metabolism in a germ-free(GF)pig model.Methods:Twelve hysterectomy-derived newborn pigs were reared in six sterile isolators.All pigs were hand-fed with sterile milk powder for 21 d,then the sterile feed was introduced to pigs for another 21 d.In the second 21-d period,six pigs were orally administrated with 25 mL/kg sterile saline per day and considered as the GF group,while the other six pigs were orally administrated with 25 mL/kg SCFAs mixture(acetic,propionic,and butyric acids,45,15,and 11 mmol/L,respectively)per day and regarded as FA group.Results:Orally administrated with SCFAs tended to increase the adiponectin concentration in serum,enhance the CPT-1 activity in longissimus dorsi,and upregulate the ANGPTL4 mRNA expression level in colon(P<0.10).Meanwhile,the mRNA abundances of ACC,FAS,and SREBP-1C in liver and CD36 in longissimus dorsi of the FA group were decreased(P<0.05)compared with those in the GF group.Besides,the mRNA expression of PGC-1αin liver and LPL in longissimus dorsi tended to(P<0.10)upregulate and downregulate respectively in the FA group.Moreover,oral administration of SCFAs tended to increase the protein level of GPR43(P<0.10)and decrease the protein level of ACC(P<0.10)in liver.Also,oral administration of SCFAs upregulated the p-AMPK/AMPK ratio and the mRNA expressions of GLUT-2 and GYS2 in liver(P<0.05).In addition,the metabolic pathway associated with the biosynthesis of unsaturated fatty acids was most significantly promoted(P<0.05)by oral administration of SCFAs.Conclusions:Exogenous introduction of SCFAs might attenuate the fat deposition and to some extent improve the

关键词： germ-free Glucose metabolism Lipid metabolism Pig model Short-chain fatty acids

Type 3 innate lymphoid cell-derived lymphotoxin prevents microbiota-dependent inflammation

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Cellular & Molecular Immunology 2018年第7期15卷 697-709页

作者： Yuan Zhang Tae-Jin Kim Joanna A Wroblewska Vera Tesic Vaibhav Upadhyay Ralph R Weichselbaum Alexei V Tumanov Hong Tang Xiaohuan Guo Haidong Tang Yang-Xin Fu Department of Pathology University of ChicagoChicagoIL 60637USA Department of Biochemistry and Molecular Biology Korea University College of MedicineSeoul 02841Korea Committee on Immunology Department of PathologyUniversity of ChicagoChicagoIL 60637USA Department of Radiation and Cellular Oncology and The Ludwig Center for Metastasis Research University of ChicagoChicagoIL 60637USA Department of Microbiology Immunology and Molecular GeneticsUniversity of Texas Health Science Center San AntonioSan AntonioTX 78229USA Chinese Academy of Sciences Key Laboratory for Infection and Immunity Institute of BiophysicsChinese Academy of SciencesBeijing 100101China Institute of Immunology Tsinghua University School of MedicineBeijing 100084China Department of Pathology University of Texas Southwestern Medical CenterDallasTX 75390USA

Splenomegaly is a well-known phenomenon typically associated with inflammation.However,the underlying cause of this phenotype has not been well characterized.Furthermore,the splenomegaly phenotype seen in lymphotoxin(LT)signaling-deficient mice is characterized by increased numbers of splenocytes and splenic neutrophils.Splenomegaly,as well as the related phenotype of increased lymphocyte counts in non-lymphoid tissues,is thought to result from the absence of secondary lymphoid tissues in LT-deficient mice.We now present evidence that mice deficient in LTα1β2 or LTβR develop splenomegaly and increased numbers of lymphocytes in non-lymphoid tissues in a microbiota-dependent manner.Antibiotic administration to LTα1β2-or LTβR-deficient mice reduces splenomegaly.Furthermore,re-derived germ-free Ltbr−/−mice do not exhibit splenomegaly or increased inflammation in non-lymphoid tissues compared to specific pathogen-free Ltbr−/−mice.By using various LTβ-and LTβR-conditional knockout mice,we demonstrate that retinoic acid-related orphan receptorγT-positive type 3 innate lymphoid cells provide the required active LT signaling to prevent the development of splenomegaly.Thus,this study demonstrates the importance of LT-mediated immune responses for the prevention of splenomegaly and systemic inflammation induced by microbiota.

关键词： germ-free lymphotoxin microbiota splenomegaly type 3 innate lymphoid cells